本文選題：深低溫保存 + 二甲基亞砜��； 參考：《山西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:本實驗用19%(v/v)的DMSO灌注兔離斷后肢,用4種不同冷凍復(fù)溫方法,對復(fù)合組織中的骨骼肌組織損傷程度進行對比研究,找到損傷程度最輕的冷凍復(fù)溫方法。方法:25只健康的新西蘭大白兔隨機分為對照組(A組)及實驗組(B、C、D、E組,B組為慢凍快復(fù)組,C組為慢凍慢復(fù)組,D組為快凍慢復(fù)組,E組為快凍快復(fù)組),實驗所用后肢均在髖關(guān)節(jié)處離斷。對照組(A組)后肢在離斷后直接用于制作標本。實驗組(B、C、D、E組)先用19%濃度DMSO(二甲基亞砜)灌注,肝素鹽水洗脫。然后B組慢速冷凍—快速復(fù)溫、C組慢速冷凍—慢速復(fù)溫、D組快速冷凍--慢速復(fù)溫、E組快速冷凍—快速復(fù)溫,然后采取與對照組相同方法制作標本,病理切片用HE染色后觀察骨骼肌損傷程度。根據(jù)病理染色的結(jié)果,對骨骼肌組織的損傷程度進行統(tǒng)計學(xué)分析。結(jié)果:經(jīng)蘇木精-伊紅(HE)染色后的骨骼肌病理切片為:對照組(A組):骨骼肌細胞核及細胞形態(tài)正常,骨骼肌組織未見肌纖維破壞及空泡形成;實驗B組:肌細胞腫脹,可見肌纖維斷裂,組織形態(tài)尚正常;實驗C組:肌細胞水腫較輕,有少量空泡,部分肌纖維斷裂;實驗D組:肌細胞水腫嚴重,少量空泡,部分肌纖維斷裂;實驗E組:肌細胞腫脹明顯,大量空泡化,肌纖維斷裂明顯,肌纖維部分溶解。結(jié)論:1.在19%(v/v)二甲基亞砜灌注條件下,實驗組(B、C、D、E組)和對照組(A組)病理切片結(jié)果之間存在顯著性差異,這表明深低溫冷凍復(fù)溫對細胞有損傷;2.各個實驗組之間的結(jié)果差異有統(tǒng)計學(xué)意義,說明骨骼肌損傷程度因冷凍復(fù)溫方式不同而不同;3.實驗組骨骼肌損傷程度由輕到重為:慢速冷凍-快速復(fù)溫組慢速冷凍-慢速復(fù)溫組快速冷凍-慢速復(fù)溫組快速冷凍-快速復(fù)溫組,可以認為在四種冷凍復(fù)溫方法中,慢速冷凍-快速復(fù)溫是最佳的冷凍復(fù)溫方法。
[Abstract]:Objective: in this experiment, 19% (v/v) DMSO was used to perfusion the hind limbs of rabbits, and 4 different cryopreservation methods were used to compare the damage degree of skeletal muscle tissue in the composite tissue, and to find the lightest degree of cryopreservation. Methods: 25 healthy New Zealand white rabbits were randomly divided into control group (group A) and experimental group (group B, C, D, E group, B group) Group C was slow freezing fast recovery group, group C was slow freezing slow recovery group, group D was fast freezing slow recovery group, group E was fast freeze-drying group, and the hind limbs were disconnected at hip joint. The hind limbs of control group (group A) were directly used to make specimens after separation. The experimental group (B, C, D, E group) was perfused with 19% concentration DMSO (two methyl sulfoxide) and heparin brine eluted. Then B group slow freezing quick freezing fast At the rapid recovery temperature, the C group slow freezing - slow rewarming, the D group fast freezing - slow rewarming, the E group fast freezing - fast rewarming, and then taking the same method as the control group to make the specimens. The pathological section was stained with HE to observe the degree of skeletal muscle injury. The damage degree of the bone muscle tissue was statistically analyzed according to the pathological staining. Results: The pathological sections of skeletal muscle after hematoxylin eosin (HE) staining were as follows: control group (group A): the nucleus and cell morphology of skeletal muscle were normal, muscle fibers were not damaged and vacuoles formed in skeletal muscle tissue; experimental B group: muscle cell swelling, muscle fiber rupture, and normal tissue morphology; experimental C group: muscle cell edema was lighter, a small amount of vacuoles, partial muscles were found. Fiber rupture; experimental group D: muscle cell edema, a small number of vacuoles, and partial muscle fiber rupture; experimental group E: muscle cells swelling obviously, a large number of vacuolation, muscle fiber fracture obvious, muscle fiber partial dissolution. Conclusion: 1. in the 19% (v/v) two methyl sulfoxide perfusion conditions, the test group (B, C, D, E group) and the control group (A group) pathological section between the results exist between the pathological section between the results exist between the pathological sections (A group) between pathological sections exist between the results of the presence of the obvious between the group (A group) pathological section between the results exist between the obvious This indicates that cryopreservation can be damaged by cryopreservation; 2. the difference between the experimental groups is statistically significant, indicating that the degree of skeletal muscle injury is different because of the different freezing methods. The degree of skeletal muscle injury in the 3. experimental group is from light to heavy: slow freezing fast rewarming group fast freezing and slow rewarming group - slow rewarming group rapid freezing rapid rewarming group, it can be considered that in the four refrigeration rewarming methods, slow freezing rapid rewarming is the best method of freezing rewarming.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R658

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